High species richness in the lichen genus Peltigera (Ascomycota, Lecanoromycetes): 34 species in the dolichorhizoid and scabrosoid clades of section Polydactylon, including 24 new to science.

Applying molecular methods to fungi establishing lichenized associations with green algae or cyanobacteria has repeatedly revealed the existence of numerous phylogenetic taxa overlooked by classical taxonomic approaches. Here, we report taxonomical conclusions based on multiple species delimitation and validation analyses performed on an eight-locus dataset that includes world-wide representatives of the dolichorhizoid and scabrosoid clades in section Polydactylon of the genus Peltigera. Following the recommendations resulting from a consensus species delimitation approach and additional species validation analysis (BPP) performed in this study, we present a total of 25 species in the dolichorhizoid clade and nine in the scabrosoid clade, including respectively 18 and six species that are new to science and formally described. Additionally, one combination and three varieties (including two new to science) are proposed in the dolichorhizoid clade. The following 24 new species are described: P. appalachiensis, P. asiatica, P. borealis, P. borinquensis, P. chabanenkoae, P. clathrata, P. elixii, P. esslingeri, P. flabellae, P. gallowayi, P. hawaiiensis, P. holtanhartwigii, P. itatiaiae, P. hokkaidoensis, P. kukwae, P. massonii, P. mikado, P. nigriventris, P. orientalis, P. rangiferina, P. sipmanii, P. stanleyensis, P. vitikainenii and P. willdenowii; the following new varieties are introduced: P. kukwae var. phyllidiata and P. truculenta var. austroscabrosa; and the following new combination is introduced: P. hymenina var. dissecta. Each species from the dolichorhizoid and scabrosoid clades is morphologically and chemically described, illustrated, and characterised with ITS sequences. Identification keys are provided for the main biogeographic regions where species from the two clades occur. Morphological and chemical characters that are commonly used for species identification in the genus Peltigera cannot be applied to unambiguously recognise most molecularly circumscribed species, due to high variation of thalli formed by individuals within a fungal species, including the presence of distinct morphs in some cases, or low interspecific variation in others. The four commonly recognised morphospecies: P. dolichorhiza, P. neopolydactyla, P. pulverulenta and P. scabrosa in the dolichorhizoid and scabrosoid clades represent species complexes spread across multiple and often phylogenetically distantly related lineages. Geographic origin of specimens is often helpful for species recognition; however, ITS sequences are frequently required for a reliable identification. Citation: Magain N, Miadlikowska J, Goffinet B, et al. 2023. High species richness in the lichen genus Peltigera (Ascomycota, Lecanoromycetes): 34 species in the dolichorhizoid and scabrosoid clades of section Polydactylon, including 24 new to science. Persoonia 51: 1-88. doi: 10.3767/persoonia.2023.51.01.

Detection of hydroquinone by Raman spectroscopy in patients with melasma before and after treatment.

BACKGROUND: Melasma is an acquired, facial hyperpigmentation without a specific origin. It is regularly associated with multiple etiologic factors such as pregnancy, genetic, racial, and from estrogen administration. Among the methods to treat skin hyperpigmentation a series of skin bleaching agents have been used. At present, the most commonly used agent is known as hydroquinone. Nowadays, it is known that hydroquinone can cause cancer in animals with unknown relevance to humans. MATERIAL AND METHODS: In this work, Raman spectroscopy was used to observe the presence of hydroquinone in the skin of 18 patients who have been under treatment for melasma. RESULTS: A significant increase in the Raman signal was observed in the six bands associated with hydroquinone after melasma treatment. CONCLUSION: The authors believe that monitoring the presence of hydroquinone may be useful for an optimal personalized treatment of melasma and to provide the specialist a support tool to control the administration of this type of bleaching agents.

Raman spectroscopy analysis of the skin of patients with melasma before standard treatment with topical corticosteroids, retinoic acid, and hydroquinone mixture.

BACKGROUND: Melasma is an abnormal acquired hyperpigmentation of the face of unknown origin, it is considered a single disease and very little has been found regarding its pathogenesis. It is usually assumed that melasma is due to excessive melanin production, but previous work using Raman spectroscopy showed degraded molecules of melanin in some melasma subjects, which may help to explain the success or failure of the standard therapy. METHODS: We perform Raman spectroscopy measurements on in vivo skin from melasma patients before treatment to identify the molecular structure of melanin within every melasma lesion. The Raman spectra were grouped according to the treatment response from patient, and the Raman spectra were analyzed. RESULTS: Raman spectroscopy measurements showed a different molecular structure of the patients who did not respond to treatment, those patients shows atypical Raman skin spectrum with peaks associated with melanin not well defined, which is consistent with molecular degradation and protein breakdown. CONCLUSION: Our results are consistent with our previous work in the sense that melasma patients who do not respond to treatment have an abnormal melanin. We believe it will eventually help to decide the treatment of melasma in clinical dermatology.

Use of Raman spectroscopy for the early detection of filaggrin-related atopic dermatitis.

BACKGROUND: Filaggrin (FLG) gene mutations, which result in complete or incomplete loss of proFLG/FLG peptides, have been reported as an important predisposing factor for atopic dermatitis (AD) and secondary atopic phenotypes such as atopic asthma. METHOD: The presence of the protein FLG in the skin was evaluated at birth on 12 infants using Raman spectroscopy; these 12 infants were monitored for 1 year to see whether they developed AD. Three different statistical analysis procedures, two of which involved principal component analysis (PCA), were performed on the Raman spectra in order to determine the FLG content. RESULTS: The infants who had a lower FLG content, determined using any of the three statistical analysis procedures proposed, were also the ones that clinically developed AD. CONCLUSION: The results suggest that Raman spectroscopy and statistical analysis such as PCA could be used as an early detection procedure for FLG -related AD and as a possible quantitative marker for FLG gene mutations.

Diffuse reflectance spectrophotometry for skin phototype determination.

BACKGROUND: The Fitzpatrick skin phototype classification scheme has become the standard method for assessing the reaction of the skin to solar stimuli; this method can be easily biased by different factors, such as ethnicity or chronic sun exposure. METHODS: Diffuse reflectance spectrophotometry (DRS) is an objective and non-invasive method used in this work to determine constitutive skin color from the upper volar arm as an objective way of measuring skin pigmentation. A DRS-determined melanin index that accounts for skin pigmentation was obtained for 35 subjects of Hispanic origin, this melanin index was compared with the physician-diagnosed and self-reported skin phototypes. RESULTS: The results show that at least for Hispanic individuals, there is a clear clinical distinction between subjects with skin phototype I and their DRS-determined melanin index; however, subjects with skin phototypes II-VI have a large melanin index overlap. CONCLUSION: Clinical assessment of skin phototype can be complemented by using DRS.

Solar elastosis and presence of mast cells as key features in the pathogenesis of melasma.

Melasma is an acquired hypermelanosis on sun-exposed areas. Its pathogenesis has not been clearly elucidated. Using histochemistry (Giemsa, Verhoeff-van Gieson and Fontana-Masson staining), we evaluated melasma lesions and compared them with nonlesional skin. Skin samples were obtained from lesional and nonlesional facial skin of 27 patients with melasma, and biopsies were also taken from normal control subjects. Mast cells and solar elastosis areas were evaluated using a computer-assisted image-analysis program. Lesional skin had abundant elastotic material compared with nonaffected skin (13.3 +/- 2.8% vs. 10.2 +/- 2.9%, P < 0.001). Mast cells were more prominent in the elastotic areas of melasma skin (173 +/- 57% vs. 145 +/- 57%, P = 0.04). Melasma could be a result of a cumulative sun exposure, in a microenvironment of cutaneous photoageing in which inflammatory cells, particularly mast cells, play a key role.

The effect of methotrexate on the expression of cell adhesion molecules and activation molecule CD69 in psoriasis.

BACKGROUND: The mechanism of the action of methotrexate (MTX) in the treatment of psoriasis has not been completely elucidated. OBJECTIVE: To assess the effect of MTX on the expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin, activation molecule CD69 and T-cell phenotype in skin specimens from patients with psoriasis. METHODS: We performed an immunohistochemical analysis of the expression of T-cell phenotype and cell adhesion/activation molecules in skin biopsies from patients with psoriasis treated with a fixed dose of MTX (12.5 mg/week). To determine data on the epidermal/dermal T-cell infiltration we carried out a manual quantification. RESULTS: Skin samples prior to therapy showed a moderate to severe inflammatory infiltrate, mainly due to T lymphocytes with a helper/inducer (CD4) phenotype. Most of these cells also expressed ICAM-1 and VCAM-1. Blood vessels showed expression of E-selectin and VCAM-1, and keratinocytes were positive for ICAM-1 staining. The cell infiltrate was reduced after therapy, as well as the expression of cell adhesion molecules. However, we also noted the persistence of the T lymphocyte phenotype CD8(+), expressing the CD69 activation molecule, after the MTX treatment. CONCLUSIONS: MTX downregulates the expression of some adhesion molecules, a phenomenon that may contribute to its anti-inflammatory therapeutic effect in psoriasis. The infiltrating T cells post-treatment have an activated cytotoxic phenotype, which may suggest a pathogenic role in the continuation and/or recurrence of psoriasis.

[Antiinflammatory effect of a topical corticosteroid: known brand versus generic drug]. / Estudio del efecto antiinflamatorio de un corticosteroide tópico: marca reconocida versus genéricos.

In Mexico, the generic drug market is growing. Regarding topical corticosteroids, there are several preparations on the local market but their clinical efficacy has not been assessed in relation with the original brand name. Using as a model the fluocinolone acetonide cream, the purpose of this study was to evaluate the antiinflammatory effect of different preparations. A double-blind, vehicle-control essay was conducted causing irritation on five sites of the volar aspect of the forearm in twenty healthy volunteers using 10% aqueous sodium lauryl sulfate. On the same part of the forearm, the formulations tested were applied for a period of 4 days with visual score irritation readings at days 2 and 4. The analysis of the results showed that although one of the generic drugs is 80% less expensive than the brandname medication, the clinical response was not as good as the other preparations. This deserves consideration, as newer and more potent generic corticosteroids will be available on the Mexican market.

Co-stimulatory signals increase the reactivity of gammadelta T cells towards mycobacterial antigens.

Although it has been shown that gammadelta T lymphocytes are able to react with different cell-associated or soluble antigens, the immune repertoire of these cells appears to be skewed to the recognition of mycobacterial antigens. We have studied the number and reactivity of gammadelta T cells towards several mycobacterial antigens in patients with tuberculosis and leprosy, as well as their healthy contacts and control individuals. We found an increased number of Vdelta2+ cells in healthy contacts (PPD+ and lepromin+) and tuberculoid leprosy patients. The gammadelta T cells from lepromatous leprosy showed a decreased response to all antigens tested, but some of these patients exhibited a significant response to the 30-kD glycoprotein of Mycobacterium tuberculosis. Interestingly, the reactivity of gammadelta T cells against mycobacterial antigens was significantly increased by costimulatory signals generated through CD7, LFA-1, CD50 and CD69 in all groups. However, signalling through CD69 did not enhance the responsiveness of gammadelta lymphocytes from lepromatous patients. On the other hand, the in vitro blockade of IL-10 with a specific antibody enhanced the cell proliferation of gammadelta lymphocytes from lepromatous leprosy patients, whereas exogenous IL-10 had an opposite effect in most individuals studied. These results suggest the potential role of different cell membrane receptors in the regulation of gammadelta T cell proliferation induced by mycobacteria, as well as the possible involvement of IL-10 in this phenomenon.

Presence of activated lymphocytes in the peripheral blood of patients with halo nevi.

BACKGROUND: The involution of the central pigmented lesion in halo nevus (HN) seems to be mediated by an immune response against self antigens expressed by melanocytes. OBJECTIVE: We assessed the presence of activated lymphocytes in the peripheral blood lymphocytes from patients with HN. METHODS: Peripheral blood was obtained from patients with HN associated with benign pigmented lesions (5) or melanoma (2) as well as from patients with melanoma without HN (5) and healthy subjects (10). Activated lymphocytes were detected by flow cytometry analysis using monoclonal antibodies (mAb) against CD69, CD71, CD98, HLA-DR, and activated beta(1) integrins (HUTS-21 mAb). RESULTS: The peripheral blood lymphocytes from patients with HN, associated with either benign or malignant lesions, exhibited a significantly higher expression of all activation markers studied compared with patients with melanoma without HN or compared with healthy subjects. Therefore the peripheral blood of HN patients contained a significant fraction of lymphocytes with an activated (CD69(+), HLA-DR(+), CD98(bright)), cell proliferating (CD71( bright)), and high adhesive (HUTS-21(bright)) phenotype. These activated cells disappeared from peripheral blood after the surgical resection of the skin lesion. CONCLUSION: Our findings further support the involvement of immune activation in HN phenomenon.

An immunohistochemical study of UV-induced skin lesions in actinic prurigo. Resistance of langerhans cells to UV light.

Actinic prurigo is an inflammatory disease of the skin that appears to be mediated by an abnormal immune response. Cell adhesion molecules play a key role in the induction of the immune response as well as in the pathogenesis of inflammation. We investigated the expression of cell adhesion and activation molecules, as well as the density of Langerhans cells in skin from patients with actinic prurigo. Skin biopsies from ultraviolet light-induced lesions, and non-irradiated areas from 10 actinic prurigo patients were studied; in addition, several spontaneous skin lesions were studied. Skin biopsies from normal individuals were used as controls. The expression of ICAM-1, ICAM-3, LFA-3, CD2, LFA-1, VLA-4, CD1a, VCAM-1, CD69, and activated b1 integrins were assessed by immunostaining. An increased expression of LFA-1, LFA-2, ICAM-3, VLA-4, and activated b1 integrins was observed in the cell infiltrate of actinic prurigo lesions and an up-regulated expression of ICAM-1 was detected in keratinocytes from these specimens. Interestingly, the number of Langerhans cells (CD1a + ) in actinic prurigo skin was not significantly affected by ultraviolet irradiation, a phenomenon that was not observed in normal controls. The increased expression of adhesion molecules in the cell infiltrate of actinic prurigo, indicates that these cells are activated and suggests that they are involved in the skin damage seen in these patients. The resistance of Langerhans cells from patients with actinic prurigo to ultraviolet light may have an important role in the pathogenesis of this condition. The involvement of keratinocytes in the pathogenesis of actinic prurigo is suggested by the expression of ICAM-1 on these cells.

Profile of cytokine mRNA expression in spontaneous and UV-induced skin lesions from actinic prurigo patients.

BACKGROUND: Actinic prurigo (AP) appears to be an immune-mediated disease triggered by exposure to ultraviolet light (UV). OBJECTIVE: To assess the profile of cytokine production in skin lesions from AP patients. METHODS: The cytokine production (IL-1 beta, IL-2, IL-4, IL-6, IL-10, IL-13, TNF-alpha, IFN-tau, and TGF-beta) in skin biopsies from 12 AP lesions was determined by a semiquantitative coupled reverse transcription-polymerase chain reaction. RESULTS: We found expression of TGF-beta and IL-13 genes in most AP skin lesions; IL-1 beta, IL-6, TNF-alpha, IFN-tau, and IL-10 were detected in some of these specimens. However, the levels of expression of all cytokines studied were not significantly different in AP skin lesions compared to nonlesional skin. CONCLUSIONS: TGF-beta and IL-13 might have a key role in both the inflammatory phenomenon and absence of significant expression of most cytokines in AP skin. The cytokine production in AP skin resembles that observed in rheumatoid synovium, a paucity in cytokine expression despite the presence of infiltrating activated mononuclear cells.

Involvement of cell adhesion and activation molecules in the pathogenesis of erythema dyschromicum perstans (ashy dermatitis). The effect of clofazimine therapy.

OBJECTIVES: To assess the expression of several cell adhesion and lymphocyte activation molecules in erythema dyschromicum perstans lesions, and to evaluate the effect of clofazimine therapy on the expression of these molecules. DESIGN AND METHODS: A prospective study. Skin biopsy samples were obtained from patients before and after 3 months of clofazimine therapy, and the expression of cell adhesion and activation molecules was assessed by an immunohistochemical technique. SETTING: This study was performed in a clinical referral center and an immunology research laboratory. PATIENTS: We studied 6 patients with erythema dyschromicum perstans. A diagnosis was made on the basis of clinical and histological criteria. Two patients discontinued participation in the study: one because of adverse effects and the other for unknown reasons. INTERVENTIONS: Patients were treated with clofazimine, 100 mg/d, for 3 months. MAIN OUTCOME MEASURES: Expression of cell adhesion and lymphocyte activation molecules in skin biopsy specimens before and after clofazimine therapy. RESULTS: Before clofazimine therapy, we detected a noticeable expression of intercellular adhesion molecule 1 and major histocompatibility complex class II molecules (HLA-DR) in the keratinocyte basal cell layer. In addition, CD36, a thrombospondin receptor that is not expressed by normal skin, was detected in the strata spinosum and granulosum. The dermal cell infiltrate expressed the activation molecule AIM/CD69 and the cytotoxic cell marker CD94. After clofazimine therapy, the expression of intercellular adhesion molecule 1 and HLA-DR disappeared, as well as the mononuclear cell infiltrate. CONCLUSIONS: Our results suggest that some cell adhesion and activation molecules are involved in the pathogenesis of erythema dyschromicum perstans. Clofazimine appears to have an important effect on the inflammatory phenomenon of erythema dyschromicum perstans.

Plasma levels and in vitro production of tumor necrosis factor-alpha and interleukin-6 in patients with amebic liver abscess.

We studied the plasma levels of tumor necrosis factor-alpha (TNF-alpha) interleukin-6 (IL-6) as well as the in vitro production of these cytokines by peripheral blood mononuclear cells, in 10 patients with amebic liver abscess (ALA) and seven healthy controls. TNF-alpha was measured using a bioassay with L929 fibroblasts; IL-6 was quantitated by the B9 cell line assay. In both assays, the number of viable cells was estimated by the conversion of MTT to formazan. TNF-alpha plasma levels were nondetectable (< 20 pg/mL) in ALA patients, as well as in the majority of healthy controls. The in vitro production of TNF induced by lipopolysaccharide was significantly decreased in ALA patients. Most ALA patients (8/10) had increased plasma levels of IL-6. Furthermore, the spontaneous production of IL-6 in vitro was significantly increased in ALA patients compared to controls. In the acute stage of the ALA, a negative relationship was found between the raised plasma levels of IL-6 and the in vitro diminished production of TNF-alpha. After recovery, ALA patients showed both normal plasma levels and in vitro production of TNF and IL-6. Our data corroborate previous reports regarding plasma levels of TNF in ALA, and suggest that E. histolytica induces the in vivo production of IL-6 through a TNF-independent pathway. The raised levels of IL-6 might in turn down-regulate the production of TNF in ALA patients.